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应用TaqMan-MGB探针进行松材线虫的实时荧光定量检测技术研究

Quantity assay of Bursaphelenchus xylophilus with TaqMan-MGB probe

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【作者】 葛建军; 曹爱新; 刘先宝; 陈长法;

【Author】 GE Jian jun CAO Ai-xin , LIU Xian-bao, CHEN Chang-fa (Chinese Academy of Quarantine and Inspection, Beijing 100029,China; South China University of Tropic Agriculture, Hainan 571737,China;Shandong Inspection and Quarantine Bureau , Qingdao 266002, China)

【机构】 中国检验检疫科学研究院; 华南热带农业大学; 山东出入境检验检疫局;

【摘要】 松材线虫是国际公认的最重要的检疫性有害生物之一,也是我国2类检疫危险性有害生物,我国口常多次从货物的木质包装中截获该线虫。由于松材线虫与拟松材线虫在形态上极其相似,难以区分,幼虫更无法用于鉴定。传统的形态学鉴定、生化以及其他分子技术等方法存在费时、准确度不高、灵敏度低等缺点,不易形成标准。我们没计筛选一对引物以及一条MGB探针,对松材线虫进行实时荧光PCR检测。建立了一条从1 pg到104pg标准曲线,相关系数r=0.965。该检测方法省时、准确、快速、无污染。

【Abstract】 Pine wood nematode (Bursaphelenchus xylophilus) , one of the most important quarantine pests in the world, is also in the A2 list concerned to China. Entry port of China intercepted it for hundreds times from wood packaging materials of goods. Due to its morphological similarity with Bursaphelenchus mucronatus, it is difficulty to distinguish it from B. mucronatus. Many attempts to develop detecting and diagnosing methods for Bursaphelenchus xylophilus had been conducted, including biochemical technology and molecular technology. However, those were time consuming and instable. A real-time quantitative polymerase chain reaction method using the MGB probe and 1 pair of primers for detecting Bursaphelenchus xylophilus was developed in this study. A liner standard curve was obtained between 1 pg and 104pg pre reaction (r =0. 965). The detection was quick, sensitive and little contamination. A faster and practicable method for detecting and diagnosis of B. xylophilus for the quarantine purpose was to be developed.

【关键词】 定量检测; 松材线虫; MGB; 核糖体ITS;
【Key words】 quantity assay; pinewood nematode; MGB; ITS;
  • 【会议录名称】 中国植物病理学会2005年学术年会暨植物病理学报创刊50周年纪念会论文集
  • 【会议名称】中国植物病理学会2005年学术年会暨植物病理学报创刊50周年纪念会
  • 【会议时间】2005-07
  • 【会议地点】中国河北保定
  • 【分类号】S763
  • 【主办单位】中国植物病理学会、中国植物病理学会病毒专业委员会
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